DETECTION OF LISTERIA MONOCYTOGENES BY THE POLYMERASE CHAIN REACTION (PCR) TECHNIQUE IN SAMPLES OF RAW BOVINE MILK
Autor(a) principal: | |
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Data de Publicação: | 2013 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | por |
Título da fonte: | Revista do Instituto de Laticínios Cândido Tostes |
Texto Completo: | https://www.revistadoilct.com.br/rilct/article/view/221 |
Resumo: | Considering the possible incidence of Listeria monocytogenes in raw foods and their pathogenicity and health risk, this study aimed to compare techniques for extraction of bacterial DNA from milk samples and investigate the presence of L. monocytogenes by Polymerase Chain Reaction (PCR) in raw milk. We tested four different extraction protocols (generally identified: A, B, C, and D) for isolation of bacterial DNA directly from milk. In all of them was obtained identifying the product of 702 bp (base pairs) corresponding to the listeriolysin gene from L. monocytogenes. The protocol B containing proteinase K and phenol buffered, was chosen for the extraction of DNA from milk samples from eight dairy farms within the RS. The subsequent PCR amplification with DNA obtained by the protocol B allowed the identification of L. monocytogenes from 103 CFU/mL. None of the samples was positive for the producer L. monocytogenes by PCR or by conventional microbiological analysis. With this study it is concluded that the tested protocols, the protocol B was more effective for the detection of L. monocytogenes by PCR. Moreover, for the samples of the producers, the result PCR technique was obtained in a shorter time than conventional analysis of L. monocytogenes, which may allow earlier treatment of infected animals and thus avoid losses to the producer. |
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DETECTION OF LISTERIA MONOCYTOGENES BY THE POLYMERASE CHAIN REACTION (PCR) TECHNIQUE IN SAMPLES OF RAW BOVINE MILKDETECÇÃO DE LISTERIA MONOCYTOGENES PELA TÉCNICA DA REAÇÃO EM CADEIA DA POLIMERASE (PCR) EM AMOSTRAS DE LEITE BOVINO IN NATURAmicro-organism; Listeriolysin; proteinase K.micro-organismo; Listeriolisina; proteinase K.Considering the possible incidence of Listeria monocytogenes in raw foods and their pathogenicity and health risk, this study aimed to compare techniques for extraction of bacterial DNA from milk samples and investigate the presence of L. monocytogenes by Polymerase Chain Reaction (PCR) in raw milk. We tested four different extraction protocols (generally identified: A, B, C, and D) for isolation of bacterial DNA directly from milk. In all of them was obtained identifying the product of 702 bp (base pairs) corresponding to the listeriolysin gene from L. monocytogenes. The protocol B containing proteinase K and phenol buffered, was chosen for the extraction of DNA from milk samples from eight dairy farms within the RS. The subsequent PCR amplification with DNA obtained by the protocol B allowed the identification of L. monocytogenes from 103 CFU/mL. None of the samples was positive for the producer L. monocytogenes by PCR or by conventional microbiological analysis. With this study it is concluded that the tested protocols, the protocol B was more effective for the detection of L. monocytogenes by PCR. Moreover, for the samples of the producers, the result PCR technique was obtained in a shorter time than conventional analysis of L. monocytogenes, which may allow earlier treatment of infected animals and thus avoid losses to the producer.Considerando a possível incidência de Listeria monocytogenes em alimentos crus e sua patogenicidade e risco para a saúde, o presente trabalho teve como objetivo comparar técnicas de extração de DNA bacteriano de amostras de leite e investigar a presença de L. monocytogenes pela Reação em Cadeia da Polimerase (PCR) em amostras de leite bovino in natura. Foram testados quatro protocolos diferentes de extração (genericamente identificados: A, B, C, e D) para o isolamento do DNA bacteriano diretamente do leite. Em todos eles foi obtida a identificação do produto de 702 pb (pares de bases) correspondente ao gene da Listeriolisina de L. monocytogenes. O protocolo B que continha proteinase K e fenol tamponado, foi o escolhido para a extração de DNA das amostras de leite de oito produtores de médio porte no interior do RS. A posterior amplificação por PCR com o DNA obtido pelo protocolo B permitiu a identificação de L. monocytogenes a partir de 103 UFC/mL. Nenhuma das amostras dos produtores foi positiva para L. monocytogenes pela técnica de PCR ou pela análise microbiológica convencional. Com o presente estudo conclui-se que, dos protocolos testados, o protocolo B foi mais eficaz para a detecção de L. monocytogenes pela técnica de PCR. Além disso, com relação às amostras dos produtores, o resultado pela técnica por PCR foi obtido em um período de tempo menor que na análise convencional de L. monocytogenes, fato que pode possibilitar um tratamento mais precoce dos animais contaminados e assim evitar perdas ao produtor.ILCTAgostini, CamilaKreling, Caroline SchwertnerBustamante-Filho, Ivan CunhaSouza, Cláucia Fernanda Volken deBiolchi, VanderleiPozzobon, Adriane2013-12-24info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://www.revistadoilct.com.br/rilct/article/view/22110.5935/2238-6416.20120073Journal of Candido Tostes Dairy Institute; v. 67, n. 389 (2012); 15-20Revista do Instituto de Laticínios Cândido Tostes; v. 67, n. 389 (2012); 15-202238-64160100-3674reponame:Revista do Instituto de Laticínios Cândido Tostesinstname:Empresa de Pesquisa Agropecuária de Minas Gerais (EPAMIG)instacron:EPAMIGporhttps://www.revistadoilct.com.br/rilct/article/view/221/231Direitos autorais 2014 Revista do Instituto de Laticínios Cândido Tostesinfo:eu-repo/semantics/openAccess2013-12-24T13:38:36Zoai:oai.rilct.emnuvens.com.br:article/221Revistahttp://www.revistadoilct.com.br/ONGhttps://www.revistadoilct.com.br/rilct/oai||revistadoilct@epamig.br|| revistadoilct@oi.com.br2238-64160100-3674opendoar:2013-12-24T13:38:36Revista do Instituto de Laticínios Cândido Tostes - Empresa de Pesquisa Agropecuária de Minas Gerais (EPAMIG)false |
dc.title.none.fl_str_mv |
DETECTION OF LISTERIA MONOCYTOGENES BY THE POLYMERASE CHAIN REACTION (PCR) TECHNIQUE IN SAMPLES OF RAW BOVINE MILK DETECÇÃO DE LISTERIA MONOCYTOGENES PELA TÉCNICA DA REAÇÃO EM CADEIA DA POLIMERASE (PCR) EM AMOSTRAS DE LEITE BOVINO IN NATURA |
title |
DETECTION OF LISTERIA MONOCYTOGENES BY THE POLYMERASE CHAIN REACTION (PCR) TECHNIQUE IN SAMPLES OF RAW BOVINE MILK |
spellingShingle |
DETECTION OF LISTERIA MONOCYTOGENES BY THE POLYMERASE CHAIN REACTION (PCR) TECHNIQUE IN SAMPLES OF RAW BOVINE MILK Agostini, Camila micro-organism; Listeriolysin; proteinase K. micro-organismo; Listeriolisina; proteinase K. |
title_short |
DETECTION OF LISTERIA MONOCYTOGENES BY THE POLYMERASE CHAIN REACTION (PCR) TECHNIQUE IN SAMPLES OF RAW BOVINE MILK |
title_full |
DETECTION OF LISTERIA MONOCYTOGENES BY THE POLYMERASE CHAIN REACTION (PCR) TECHNIQUE IN SAMPLES OF RAW BOVINE MILK |
title_fullStr |
DETECTION OF LISTERIA MONOCYTOGENES BY THE POLYMERASE CHAIN REACTION (PCR) TECHNIQUE IN SAMPLES OF RAW BOVINE MILK |
title_full_unstemmed |
DETECTION OF LISTERIA MONOCYTOGENES BY THE POLYMERASE CHAIN REACTION (PCR) TECHNIQUE IN SAMPLES OF RAW BOVINE MILK |
title_sort |
DETECTION OF LISTERIA MONOCYTOGENES BY THE POLYMERASE CHAIN REACTION (PCR) TECHNIQUE IN SAMPLES OF RAW BOVINE MILK |
author |
Agostini, Camila |
author_facet |
Agostini, Camila Kreling, Caroline Schwertner Bustamante-Filho, Ivan Cunha Souza, Cláucia Fernanda Volken de Biolchi, Vanderlei Pozzobon, Adriane |
author_role |
author |
author2 |
Kreling, Caroline Schwertner Bustamante-Filho, Ivan Cunha Souza, Cláucia Fernanda Volken de Biolchi, Vanderlei Pozzobon, Adriane |
author2_role |
author author author author author |
dc.contributor.none.fl_str_mv |
|
dc.contributor.author.fl_str_mv |
Agostini, Camila Kreling, Caroline Schwertner Bustamante-Filho, Ivan Cunha Souza, Cláucia Fernanda Volken de Biolchi, Vanderlei Pozzobon, Adriane |
dc.subject.none.fl_str_mv |
|
dc.subject.por.fl_str_mv |
micro-organism; Listeriolysin; proteinase K. micro-organismo; Listeriolisina; proteinase K. |
topic |
micro-organism; Listeriolysin; proteinase K. micro-organismo; Listeriolisina; proteinase K. |
description |
Considering the possible incidence of Listeria monocytogenes in raw foods and their pathogenicity and health risk, this study aimed to compare techniques for extraction of bacterial DNA from milk samples and investigate the presence of L. monocytogenes by Polymerase Chain Reaction (PCR) in raw milk. We tested four different extraction protocols (generally identified: A, B, C, and D) for isolation of bacterial DNA directly from milk. In all of them was obtained identifying the product of 702 bp (base pairs) corresponding to the listeriolysin gene from L. monocytogenes. The protocol B containing proteinase K and phenol buffered, was chosen for the extraction of DNA from milk samples from eight dairy farms within the RS. The subsequent PCR amplification with DNA obtained by the protocol B allowed the identification of L. monocytogenes from 103 CFU/mL. None of the samples was positive for the producer L. monocytogenes by PCR or by conventional microbiological analysis. With this study it is concluded that the tested protocols, the protocol B was more effective for the detection of L. monocytogenes by PCR. Moreover, for the samples of the producers, the result PCR technique was obtained in a shorter time than conventional analysis of L. monocytogenes, which may allow earlier treatment of infected animals and thus avoid losses to the producer. |
publishDate |
2013 |
dc.date.none.fl_str_mv |
2013-12-24 |
dc.type.none.fl_str_mv |
|
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://www.revistadoilct.com.br/rilct/article/view/221 10.5935/2238-6416.20120073 |
url |
https://www.revistadoilct.com.br/rilct/article/view/221 |
identifier_str_mv |
10.5935/2238-6416.20120073 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.none.fl_str_mv |
https://www.revistadoilct.com.br/rilct/article/view/221/231 |
dc.rights.driver.fl_str_mv |
Direitos autorais 2014 Revista do Instituto de Laticínios Cândido Tostes info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Direitos autorais 2014 Revista do Instituto de Laticínios Cândido Tostes |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
ILCT |
publisher.none.fl_str_mv |
ILCT |
dc.source.none.fl_str_mv |
Journal of Candido Tostes Dairy Institute; v. 67, n. 389 (2012); 15-20 Revista do Instituto de Laticínios Cândido Tostes; v. 67, n. 389 (2012); 15-20 2238-6416 0100-3674 reponame:Revista do Instituto de Laticínios Cândido Tostes instname:Empresa de Pesquisa Agropecuária de Minas Gerais (EPAMIG) instacron:EPAMIG |
instname_str |
Empresa de Pesquisa Agropecuária de Minas Gerais (EPAMIG) |
instacron_str |
EPAMIG |
institution |
EPAMIG |
reponame_str |
Revista do Instituto de Laticínios Cândido Tostes |
collection |
Revista do Instituto de Laticínios Cândido Tostes |
repository.name.fl_str_mv |
Revista do Instituto de Laticínios Cândido Tostes - Empresa de Pesquisa Agropecuária de Minas Gerais (EPAMIG) |
repository.mail.fl_str_mv |
||revistadoilct@epamig.br|| revistadoilct@oi.com.br |
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1809738130914279425 |